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[噬菌体T4的DNA BglII片段在载体质粒pSCC31中的克隆]

[Cloning of the DNA BglII fragments of bacteriophage T4 in the vector plasmid pSCC31].

作者信息

Nivinskas R G

出版信息

Genetika. 1988 Jan;24(1):34-41.

PMID:2833423
Abstract

An attempt has been made to clone six BglII fragments of T4 DNA in the range of 3.3-8.1 kb in the vector plasmid pSCC31 containing a single BglII site within the gene for endonuclease EcoRI and pL promoter of phage lambda. DNA fragments were extracted from the corresponding bands of agarose gel. The following BglII fragments were cloned: the 3.3 kb fragment No. 9 containing a portion of gene 20, the gene 21 and a portion of gene 22; the 4.2 kb fragment No. 8.1 with genes 17, 18, 19 and a portion of gene 20; the 5.2 kb fragment No. 7.1 with genes 25-29 and a portion of gene 48. In the case of the fragment No. 7.1, the recombinant plasmids pRL705 and pRL707 with different orientation of phage DNA fragment were obtained. An attempt to clone the fragments No. 8.2 (4.2 kb), No. 7.2 (5.45 kb) and No. 6 (8.1 kb) was unsuccessful and this probably indicates the presence of the genes, whose products are deleterious to the growth of bacterial cell.

摘要

已尝试将T4 DNA的6个BglII片段(大小在3.3 - 8.1 kb之间)克隆到载体质粒pSCC31中,该质粒在噬菌体λ的内切酶EcoRI基因和pL启动子内含有一个单一的BglII位点。从琼脂糖凝胶的相应条带中提取DNA片段。克隆了以下BglII片段:含有基因20一部分、基因21和基因22一部分的3.3 kb片段9;含有基因17、18、19和基因20一部分的4.2 kb片段8.1;含有基因25 - 29和基因48一部分的5.2 kb片段7.1。对于片段7.1,获得了噬菌体DNA片段方向不同的重组质粒pRL705和pRL707。克隆片段8.2(4.2 kb)、7.2(5.45 kb)和6(8.1 kb)的尝试未成功,这可能表明存在其产物对细菌细胞生长有害的基因。

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